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OfflineDocShamen
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agar firsr time
    #23727039 - 10/10/16 10:57 PM (7 years, 3 months ago)

I've poured my agar to plates i took tissue from within the stem. im not seeing strong white strands rather i can barely notice them until i put them under a light then i can see the growth.


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Offlinetump
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Re: agar firsr time [Re: DocShamen]
    #23727046 - 10/10/16 11:00 PM (7 years, 3 months ago)

What was the nuke % count. Matted growth is normal


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Re: agar firsr time [Re: tump]
    #23727092 - 10/10/16 11:20 PM (7 years, 3 months ago)

Pic's would be good.


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Re: agar firsr time [Re: Pastywhyte]
    #23727109 - 10/10/16 11:28 PM (7 years, 3 months ago)

:whathesaid:


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OfflineDocShamen
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Re: agar firsr time [Re: Pastywhyte]
    #23727127 - 10/10/16 11:37 PM (7 years, 3 months ago)

lol like i said you can barely see it but i did the best i could


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Re: agar firsr time [Re: DocShamen]
    #23727137 - 10/10/16 11:42 PM (7 years, 3 months ago)

Wrap has to go all the way around the plate

Like these

Sometimes growth can be clearish and wispy


Edited by bodhisatta (10/10/16 11:43 PM)


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OfflineDocShamen
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Re: agar fi rsr time [Re: bodhisatta]
    #23727140 - 10/10/16 11:47 PM (7 years, 3 months ago)

the last plate more like what i have. but mine is harder to see without angling it to the light or holding a flashlight underneath
edit: also whats the reasoning for wrap all the way around besides for keeping contams out?


Edited by DocShamen (10/10/16 11:49 PM)


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Re: agar fi rsr time [Re: DocShamen]
    #23727147 - 10/10/16 11:49 PM (7 years, 3 months ago)

Keeping contamination out.

You would be surprised where ants and fruit flies end up


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Re: agar fi rsr time [Re: DocShamen]
    #23727148 - 10/10/16 11:49 PM (7 years, 3 months ago)

What type of nutes did you use? Nutrition profile is either too low or the wrong kind.


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OfflineDocShamen
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Re: agar fi rsr time [Re: Pastywhyte]
    #23727152 - 10/10/16 11:53 PM (7 years, 3 months ago)

got some premixed PDA


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Re: agar fi rsr time [Re: DocShamen]
    #23727157 - 10/10/16 11:57 PM (7 years, 3 months ago)

How much did you use? It looks like starved agar to me.


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OfflineDocShamen
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Re: agar fi rsr time [Re: DocShamen]
    #23727162 - 10/11/16 12:01 AM (7 years, 3 months ago)

10g/250ml


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Re: agar fi rsr time [Re: DocShamen]
    #23727165 - 10/11/16 12:03 AM (7 years, 3 months ago)

Well that should work but obviously isn't. Is this the first time you used this agar? Has it worked before? You taste it at all?


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Re: agar fi rsr time [Re: Pastywhyte]
    #23727173 - 10/11/16 12:06 AM (7 years, 3 months ago)

Premade pda is weird sometimes. Saccharomyces cerevisiae grows just fine but sometimes cube mycelium acts strange. Also most premade media is 12-15g/L agar rather than 20g/L like we use for most fungi with average gel strength agar(seems most is about the same)

Premade YPD is largely for yeast and some specific bacteriological work
Things that grow in way more aqueous environments usually. Also they incubate dishes which dries out dishes and they incubate upside down so the condensation goes on the lid. Then the lid is tossed out and the plate isn't saved usually.

Hence 12-15g/L having poor performance usually. Look at your mix and post the actual recipe. You may have to fortify the media with some more agar agar to get to 2% agar or 20g/L(with 1L water=1000g for all intensive purposes)

Mycelium growth is better on this firm media. And we don't dry our plates first because we're trying to be aseptic as possible. Liquidy agar is fine for germination of spores but you may want to firm it up to try to get growth that's not so clear. It's basically drowning on the surface is my thoughts and only getting o2 through diffusion through the thin layer of water hence shit performance.


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Re: agar fi rsr time [Re: Pastywhyte]
    #23727176 - 10/11/16 12:08 AM (7 years, 3 months ago)

Quote:

Pastywhyte said:
Well that should work but obviously isn't. Is this the first time you used this agar? Has it worked before? You taste it at all?



Test*

Hahaha man I was confused as hell for a while


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Re: agar fi rsr time [Re: bodhisatta]
    #23727184 - 10/11/16 12:11 AM (7 years, 3 months ago)

Man I have never had any issue with potato based agar ever, it's more reliable than malt for me. But then again I only make mine myself and if some of the ratios were low that would totally explain the look of that culture.

*Fucking autocorrect.


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OfflineDocShamen
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Re: agar fi rsr time [Re: Pastywhyte]
    #23727188 - 10/11/16 12:13 AM (7 years, 3 months ago)

first time using it yes. havent tasted it. on some plates it will look clear strands like that but with more white at the tips. in some areas.
next time i may use 20g/250ml. agar does seem more clear than other peoples agar.


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Re: agar fi rsr time [Re: bodhisatta]
    #23727193 - 10/11/16 12:14 AM (7 years, 3 months ago)

Heres what I found for that exact same package of agar in the picture. Here's the description
Quote:



Back
Description
100 grams Potato Dextrose Agar (Dehydrated) PDA FORMULA: 24% Dried Potato Powder 43% Agar Agar Powder 33% Dextrose Powder 100 grams of PDA is sufficient to create a 2500ml solution which will yield 80-90 100mm petri dishes Potato dextrose agar is a common microbiological growth media made from potato infusion, and dextrose. Potato dextrose agar (abbreviated "PDA") is the most widely used medium for growing fungi and bacteria which attack living plants or decaying dead plant matter.






So 43g in 100g of total. So that's 43g agar in 2500ml water
So that's 17.2g in 1000ml or 1.7% I would think that would work but use more powder and less water 2100-2200mL to a 100g package

Or for 20g use 420-440mL
And 10g use 210-220ml


Seems oddly marginal for that big of growth difference but ive seen weirder.


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OfflineDocShamen
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Re: agar fi rsr time [Re: bodhisatta]
    #23727209 - 10/11/16 12:20 AM (7 years, 3 months ago)

if i used 20g/250ml just doubling the pda than whats on the container, would that be to much nutes. would it cause problems in growth?


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Re: agar fi rsr time [Re: DocShamen]
    #23727227 - 10/11/16 12:28 AM (7 years, 3 months ago)

Try it out. It's 24g in 2500mL so .96% potato basically 1%
And 33g glucose to 2500mL is 1.3%

I think you could double it no problem.

It's 2500mL so to find these percentages with no calculator think like
2500*4=10,something zeros don't matter
Then multiply 24g or 33g by 4 so
96 and 132 so you have your percentages now. .96 and 1.32


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OfflineDocShamen
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Re: agar fi rsr time [Re: bodhisatta]
    #23727242 - 10/11/16 12:34 AM (7 years, 3 months ago)

thank you for both of your response and i will keep this updated. ill try it out on some wide mouth 1/2 pints sense im out of plates atm.
do you think ill be able to get a cut from those plates and hopefully colonize?


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Re: agar fi rsr time [Re: DocShamen]
    #23727398 - 10/11/16 04:50 AM (7 years, 3 months ago)

I was having weak growth on my pre made pda agar to find out it was 33 percent dextrose.


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Re: agar fi rsr time [Re: Greenskybluegrass7]
    #23727506 - 10/11/16 06:46 AM (7 years, 3 months ago)

If you transfer from the low nute plates it should get really aggressive. I would give it a try.


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Re: agar fi rsr time [Re: Greenskybluegrass7]
    #23727714 - 10/11/16 09:00 AM (7 years, 3 months ago)

Quote:

Greenskybluegrass7 said:
I was having weak growth on my pre made pda agar to find out it was 33 percent dextrose.



Did you see my post om the other page. The pda is 33% dextrose but you use 100g to 2500ML.

So that's only 33g dextrose to 2500mL
So that's only 1.32% dextrose actually


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